Cell Applications, Inc.

This is our hallmark service: We isolate, expand, freeze, document, and test cells from the tissue of your choice, supplied by you or acquired by CAI. We offer a number of additional cell-based services and assays, indicated below.

Our scientists hold deep experience in primary cell purification. Cells are carefully removed and released from tissue, and then established as a population in a controlled culture environment. The primary cells in our comprehensive library are low passage and robust, with high viability, transfection efficiency and guaranteed doublings. To maintain optimal cell health, behavior and performance, our media, reagents and conditions are carefully optimized for each cell type. We can expand, freeze, document and test cells isolated from nearly any tissue or donor profile

Components & supplements are carefully formulated to the unique requirements of the specific cell type. This optimization enhances cell health, viability, population doublings, physiology, behavior, sub-culture, data integrity and reproducibility. Serum- & Xeno-free & other versions available. Confirmed free of bacterial, yeast and fungal contamination.

CAI media are tested for sterility in order to confirm no bacteria, yeast or fungi contaminate the solutions. The products undergo further quality control for correct pH, osmolality and lack of endotoxins. A panel of different bioassays affirm the media sustain a proper environment for expected cell-type-specific culture, growth, plating, karyotype, physiology, morphology, viability, population doublings, surface markers, cryopreservation, differentiation and/or induction.

Media are carefully optimized for the characteristics and requirements unique to each cell type. Attention to detail ensures ideal cell health, viability, performance, physiology, morphology, consistency and data.

Primary cells and paired media have been optimized for pharmaceutical drug discovery efforts addressing genetic and autoimmune disorders, injury, inflammation and more. These cellular disease models include Asthma, Chronic Obstructive Pulmonary Disease (COPD), Osteoarthritis (OA), Rheumatoid Arthritis (RA) and Type II Diabetes (T2D). We are also happy to prepare custom disease cell panels for your specific screening requirements.

Our 3D Cell Models include Airway, with Human Bronchial Epithelial Cells differentiated into a pseudostriated epithelium, and Skin, where Human Epidermal Keratinocytes are differentiated into stratified squamous epithelium. Both serve as highly physiological three dimensional tissue models for in vitro studies, providing excellent cellular systems to study epithelial function and disease. The models, created by growing cells on inserts with a liquid/air interface and specialized culture conditions, are offered pre-formed or as a kit containing all necessary materials and detailed instructions.

Equipment: Cyfuse Biomedical Regenova Bio 3D Printer
Reagents: Primary Cells, Media
Software: Special B3D software for designing three dimensional tissue structure
Species: Human & Animal

Advanced tissue-engineering services are available using a new three-dimensional bioprinting approach called the “Kenzan Method”. Utilizing Cyfuse Biomedical’s Regenova® 3D Bio Printer, a robotic system that fabricates three-dimensional tissue from primary cells, Cell Applications has created a powerful bio-printing model that makes scaffold-free, primary cell-based tissue available.

The instrument assembles 3D macroscopic tissue by lancing primary cell spheroids onto a fine needle array. Pre-programmed software guides the collection of the spheroids, which can then be designed and fused into various tissue configurations. As the cells mature, they self-organize, promoting strong, reliable tissue that is further optimized by the design of the bio printer’s needle array that allows for optimum circulation of culture medium. The Method is a gentler approach and increases the percentage of viable cells. The method has already been utilized or assessed to create blood vessels, nerve and functional liver-like tissues. Other anticipated applications include cardiac muscle and pancreatic islets, as well as spinal cord, urologic, trachea, skin and digestive tissues.

Customers can select from CAI's extensive bank of standard human and animal primary cells. Alternatively, primary cells can be custom-isolated from virtually and mammalian tissue, whether normal or diseased.

INTEGRATION-FREE IPSC GENERATION AND VALIDATION

Our HiPSC are generated by using the RNA-based Sendai virus to deliver reprogramming factors to donor skin fibroblasts. Since the Sendai virus is an RNA virus that does not go through a DNA phase, its genetic material as well as the transgenes do not integrate into the host cells genome.

Reprogramming triggers a cascade of evident changes in the host cells that are recognizable morphologically and through a combination of markers and pluripotency assays. Our HiPSCs display classic pluripotent stem cell morphology, with a high nucleus to cytoplasm size ratio, as well as they are amenable to be cultivated in serum-free media, independent of feeder cells and of feeder-conditioned media as colonies or high density monolayers. hiPSCs are also evaluated for the presence of karyotypic abnormalities. Confirmation of pluripotency is performed through the analysis of expression of several established independent pluripotency markers. Unguided differentiation confirms HiPSC ability to generate cell derivatives of tissues arising from the three embryonic layers.
Cell Characterization. Post-thawing viability of HiPSCs is typically higher than 70%, and HiPSC have demonstrated coherent pluripotent behavior over more than 60 passages. Although it is in theory possible to propagate HiPSCs indefinitely, HiPSC subculturing over passages higher than is usually not recommended, as the chances of karyotypic abnormalities increase. HiPSCs are also tested to ensure absence of microorganism contaminants.

DOCUMENTED CONSISTENCY
Uniform
Reproducible
Chemically defined growth conditions
Detailed, specified expansion protocols
Comprehensive documentation

QUALITY CONTROL & TESTING
Pluripotency assays
Imaging & Morphology
Karyotype
Documentation
Post-thaw viability

PRODUCT SELECTION & SUPPORT
Cryopreserved in aliquots of 1 Million viable cells
Technical support and product guidance
Experienced PhD scientists available
Specialized, optimized media

IPSC CUSTOM SERVICES
Available on demand
Cell expansion
Disease models
Scale-able
Need 100 million to >1B cells? Call Us!

Custom Differentiation of Human Induced Pluripotent Stem Cells (HiPSC) into into cell derivatives from the 3 embryonic layers: Neuronal, Smooth Muscle and Hepatic.

"More time for research. Less time preparing cells"

IPSC DIFFERENTIATION SERVICES
On-demand production
HiPSC expansion
Differentiation of iPSC-derived cells
Disease models
Multi-Cell Differentiation
Reproducible differentiation
Many lines in parallel
Multiple mutations from single donor
Single cell type from multiple donors
Efficient
Fast expansion time
Resource-friendly
Fewer steps
Higher productivity
Unprecedented Scale
Massively parallel production
Large quantities of iPSC-derived cells

1 Billion HiPSC per lot expanded in a week
Cost-efficient, modular, scale-able protocol

Contact Us to Start Your Custom iPSC Project

IPSC MEDIUM OVERVIEW
Quality-tested growth & freezing media, coating solution and subculture reagents, carefully optimized for the characteristics and requirements unique to Human Induced Pluripotent Stem Cells (HiPSC). Attention to detail ensures ideal cell health, viability, performance, physiology, morphology, consistency and data.

FOR USE WITH
Human Induced Pluripotent Stem Cells:HiPSC

IPSC MEDIA & REAGENTS from Cell Applications, Inc.

• HiPSC Growth Medium: Xeno-free, chemically-defined medium used to thaw, culture and propagate HiPSC, supporting growth as a monolayer (analogous to Essential 8TM - E8), Use at room temperature

• HiPSC Freezing Medium: Serum-free cryopreservation formula, containing sucrose & dextran

• HiPSC Subculture Reagent (Dissociation Medium): Ca2+ & Mg2+ - free formulation containg metal ion chelator ethylenediaminetetraacetic acid in buffered saline

• HiPSC Coating Solution: Pre-diluted, ready-for-use preparation of growth factor-reduced basement membrane components, extracted from Mouse Englebreth Holm Swarm (EHS) tumor (analogous to Matrigel®)

• ROCK Inhibitor: Y-27632: A ROCK1-selective, small molecule Rho-Associated Protein Kinase Inhibitor known to increase stem cell survival & cloning efficiency, without affecting pluripotency

Highly sensitive β-Gal Assay Kits provide convenient, accurate quantitation of beta galactosidase in cultured cells, down to the lowest expression levels .

Cell signaling pathway strength varies, requiring valuable resources to screen cells for sensitivity to common stimuli. Save time and do better science with pre-screened cells.

• Endothelial Cells: Aortic, Umbilical Vein & Microvascular ECs stimulated by VEGF, assayed for activation of VEGFR2 signaling pathway, with others custom pre-screened by special request
• Skeletal Muscle Cells stimulated with insulin & oligomycin, verified for activation of Akt, MAPK, AMPK & GSK cell signaling
  *Preadipocytes differentiated into Adipocytes, assayed for adipocyte & metabolic regulation markers, e.g. PPAR, CEBP, Insulin Receptor & Adiponectin

Cytofect™ Transfection Kits contain a new class of unique chemical reagent formulations specifically optimized for transfecting primary cells, all in one complete, affordable system. Researchers routinely cite Cytofect's high efficiency and low toxicity.

We can prepare Total RNA and/or Poly A+RNA from the cell types or tissue you specify. The highly pure RNA is ready for downstream applications like microarray hybridization, PCR cDNA synthesis and RT-PCR analysis. Quality control tests confirm RNA integrity and stability, with no genomic DNA contamination.

Cyto-X: High sensitivity and cell viability. Cyto-X is a ready-for-use cell viability reagent for proliferation assays. When added to cell culture, a highly water-soluble tetrazolium salt is reduced by dehydrogenases in live cells to form an orange colored formazan product that is soluble in culture medium. The amount of formazan product is directly proportional to the enzymatic activity of dehydrogenases and the number of living cells. With its high sensitivity and low cytotoxicity, Cyto-X outperforms traditional assays using other tetrazolium salts such as MTT, XTT, MTS or WST-1.

Cyto-X: High sensitivity and cell viability. Cyto-X is a ready-for-use cell viability reagent for proliferation and cytotoxicity assays. When added to cell culture, a highly water-soluble tetrazolium salt is reduced by dehydrogenases in live cells to form an orange colored formazan product that is soluble in culture medium. The amount of formazan product is directly proportional to the enzymatic activity of dehydrogenases and the number of living cells. With its high sensitivity and low cytotoxicity, Cyto-X outperforms traditional assays using other tetrazolium salts such as MTT, XTT, MTS or WST-1.